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MIQE: Real-Time PCR Experiment Standards

The potential applications for Quantitative Real-Time PCR (qPCR) have increased exponentially since the first description (Higuchi, 1993). However, researchers have been frustrated by complications such as contamination, insufficient amplification, low sensitivity, and uncertainty about what constitutes a suitable statistical analysis. Until recently, there has been a lack of consensus about how to deal with these obstacles.

An international research team, including Dr. Tania Nolan, our Global Manager for Applications and Technical Support, published The MIQE (pronounced Mykee) Guidelines in 2009 to address the challenges of performing dependable qPCR measurements. By following MIQE, you are certain to produce more reliable data and will:

  • Promote experimental transparency
  • Ensure consistency between laboratories
  • Maintain the integrity of the scientific literature

Our qPCR services, including primer and probe designs, assay protocol development,  troubleshooting, and data analysis support, adhere to MIQE, which allows you to publish or bring your product to market faster and with confidence.

The MIQE Checklist Quick Reference Guide

1Clinical chemistry Copyright 2009 by AMERICAN ASSOCIATION FOR CLINICAL CHEMISTRY, INC. Reproduced with permission of AMERICAN ASSOCIATION FOR CLINICAL CHEMISTRY, INC in the format Internet posting via Copyright Clearance Center.

2All essential information must be submitted with the manuscript. Desirable information should be submitted if available. If primers are from RTPrimerDB, information on qPCR target, oligonucleotides, protocols, and validation is available from that source.

3Assessing the absence of DNA using a no-reverse transcription assay is essential when first extracting RNA. Once the sample has been validated as DNA-free, inclusion of no-reverse transcription control is desirable but no longer essential.

4Disclosure of the probe sequence is highly desirable and strongly encouraged. However, because not all commercial pre-designed assay vendors provide this information, it cannot be an essential requirement. Use of such assays is discouraged.

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